NUCLEOPROTEIN-BASED ELISA FOR DETECTION OF SARS-CoV-2 IgG ANTIBODIES: COULD AN OLD ASSAY BE SUITABLE FOR SERODIAGNOSIS OF THE NEW CORONAVIRUS?

Tozetto-Mendoza, Tania Regina; Kanunfre, Kelly Aparecida; Vilas-Boas, Lucy Santos; Sanchez Espinoza, Evelyn Patricia; Paião, Heuder Gustavo Oliveira; Rocha, Mussya Cisotto; de Paula, Anderson Vicente; de Oliveira, Maura Salaroli; Zampelli, Daniella Bosco; Vieira, José Mauro; Buss, Lewis; Costa, Silvia Figueiredo; Sabino, Ester Cerdeira; Witkin, Steven S.; Okay, Thelma Suely; Mendes-Correa, Maria Cassia

OBJECTIVES: We evaluated the performance of a nucleoprotein-based enzyme-linked immunosorbent assay (ELISA) for detection of IgG antibodies to SARS-CoV-2. METHODS: The ELISA was based on serum IgG reactivity to a 46-kDa protein derived from the recombinant SARS-CoV2 nucleoprotein. Assay sensitivity was assessed using serum samples from 134 COVID-19 confirmed cases obtained > 15 days after symptom onset. Specificity was determined by testing sera from 94 healthy controls. Cross-reactivity was evaluated with sera from 96 individuals with previous dengue or zika virus-confirmed infections, with 44 sera from individuals with confirmed infections to other respiratory viruses or with bacterial and fungal infections that cause pneumonia and with 40 sera negative for SARS-CoV-2 nucleoprotein by commercial ELISA kits. RESULTS: The majority of subjects were male and> 60 years old. Assay sensitivity was 90.3% (95% confidence interval 84.1% - 94.2%) and specificity was 97.9% (92.6% - 99.4%). There was no cross-reactivity with sera from individuals diagnosed with dengue, zika virus, influenza virus, rhinovirus, adenovirus, respiratory syncytial virus, seasonal coronavirus, Mycobacterium tuberculosis, Staphylococcus (S. aureus and coagulase-negative), Streptococcus pneumoniae, Klebsiella pneumoniae and the fungus Aspergillus fumigatus. The level of concordance of our test with results from commercial ELISA kits was 100%. CONCLUSION: The nucleoprotein-based ELISA was specific for detection of IgG anti-nucleoprotein antibodies to SARS-CoV-2.. It utilizes a frequently employed low expense assay protocol and is easier to perform than other currently available commercial SARS-CoV2 antibody detection tests.